Xanthan Gum Capped Zno Microstars As a Promising Dietary Zinc Supplementation
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foods Article Xanthan Gum Capped ZnO Microstars as a Promising Dietary Zinc Supplementation Alireza Ebrahiminezhad 1,2, Fatemeh Moeeni 2, Seyedeh-Masoumeh Taghizadeh 2, Mostafa Seifan 3, Christine Bautista 3, Donya Novin 3, Younes Ghasemi 2,* and Aydin Berenjian 3,* 1 Department of Medical Nanotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz 71348, Iran; [email protected] 2 Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz 71348, Iran; [email protected] (F.M.); [email protected] (S.-M.T.) 3 School of Engineering, Faculty of Sciences and Engineering, University of Waikato, Hamilton 3216, New Zealand; [email protected] (M.S.); [email protected] (C.B.); [email protected] (D.N.) * Correspondence: [email protected] (Y.G.); [email protected] (A.B.) Received: 7 February 2019; Accepted: 26 February 2019; Published: 2 March 2019 Abstract: Zinc is one of the essential trace elements, and plays an important role in human health. Severe zinc deficiency can negatively affect organs such as the epidermal, immune, central nervous, gastrointestinal, skeletal, and reproductive systems. In this study, we offered a novel biocompatible xanthan gum capped zinc oxide (ZnO) microstar as a potential dietary zinc supplementation for food fortification. Xanthan gum (XG) is a commercially important extracellular polysaccharide that is widely used in diverse fields such as the food, cosmetic, and pharmaceutical industries, due to its nontoxic and biocompatible properties. In this work, for the first time, we reported a green procedure for the synthesis of ZnO microstars using XG, as the stabilizing agent, without using any synthetic or toxic reagent. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM) were used to study the structure, morphology, and size of the synthesized ZnO structures. The results showed that the synthesized structures were both hexagonal phase and starlike, with an average particle size of 358 nm. The effect of different dosages of XG-capped ZnO nanoparticles (1–9 mM) against Gram-negative (Escherichia coli) and Gram-positive (Bacillus licheniformis, Bacillus subtilis, and Bacillus sphaericus) bacteria were also investigated. Based on the results, the fabricated XG-capped ZnO microstars showed a high level of biocompatibility with no antimicrobial effect against the tested microorganisms. The data suggested the potential of newly produced ZnO microstructures for a range of applications in dietary supplementation and food fortification. Keywords: xanthan gum; ZnO microparticles; ZnO microstars; bacteria; supplementation; food fortification 1. Introduction Zinc plays many essential roles in all life forms [1], and is part of the major subgroup of micronutrients that are beneficial in human nutrition and health [2]. In the human diet, the main sources of zinc are animal products [3], rice, wheat, and soybean [4]. However, there has been an insufficient dietary supply of the element, which has become the primary cause of zinc deficiency. Studies have revealed that the concentrations of zinc mineral elements in edible parts have decreased over the last 50 years [5], and that high levels of dietary inhibitors may suppress the absorption of Foods 2019, 8, 88; doi:10.3390/foods8030088 www.mdpi.com/journal/foods Foods 2019, 8, 88 2 of 10 zinc [6]. Zinc deficiency has an effect on organ systems such as the epidermal, gastrointestinal, central nervous, immune, skeletal, and reproductive systems [7]. The literature has also determined that zinc deficiency can impair physical growth, and zinc-dependent metabolic functions [2]. It is essential for an individual to consume a certain level of zinc, as the zinc deficiency can have several consequences on human health. There are currently four main approaches to increasing the intake of zinc in the human diet: (1) direct supplementation, (2) food fortification, (3) dietary diversification, and (4) crop biofortification [5]. To compensate for insufficient zinc intake, the major suggested route is through the intake of oral zinc supplementation. The forms of zinc that are frequently used in supplements are zinc sulfate, zinc oxide, zinc acetate, and zinc gluconate [8]. However, zinc supplements are not well tolerated by all patients, even in high doses, due to inadequate intestinal zinc absorption [6]. Currently, there is great attention on the use of micro and nanoparticles to fortify micronutrients, due to their ultra-small size which aids intestinal absorption. Micronization is another effective technique to increase the digestive absorption of poorly soluble nutrients and drugs. It has been reported that the decrease in the particle size, up to a few microns, can substantially enhance the rate of dissolution and can consequently improve the bioavailability and clinical efficacy [9]. Although, in contrast to metal salts, which are commonly prescribed, metal and metal oxide particles are more tolerable for patients, and therefore, higher dosages can be used [6]. Nanometer-sized particles of zinc oxide (ZnO) have many applications in various areas due to their unique and superior chemical and physical properties compared to bulk ZnO. Over the last decade, massive investigations have also been performed to evaluate the effect of ZnO nanoparticles on the growth of microbial communities. For example, Ramesh et al. [10] investigated the antibacterial activity of green-synthesized ZnO nanoparticles with an average diameter of 29.79 nm. The zone inhibition method was used to explore the antibacterial activity of ZnO nanoparticles against Gram-positive (Staphyloccus aureus) strains, and Gram-negative (Salmonella paratyphi, Vibrio cholerae, Escherichia coli) strains. Authors have concluded that the ZnO nanoparticles exhibit good antibacterial activities by decreasing the cell growth. Banoee et al. [11] conducted a similar study which explored the antibacterial activity of ZnO nanoparticles with an average size of 20–45 nm against S. aureus and E. coli. It was observed that the presence of fabricated ZnO nanoparticles enhanced the antibacterial activity of ciprofloxacin, and increased the inhibition zone by 27% and 22%, for S. aureus and E. coli, respectively. However, the availability of 500 µg/disk ZnO nanoparticles decreased the antibacterial activity of amoxicillin, penicillin G, and nitrofurantoin in S. aureus. The antibacterial behavior of Vitex negundo extract assisted ZnO nanoparticles against S. aureus (ATCC 11632) and E. coli (ATCC 10536), and was studied by Ambika et al. [12] using the agar well diffusion method. The results showed that synthesized ZnO nanoparticles have high antibacterial activity. It was shown that the availability of ZnO nanoparticles contributed to a zone of inhibition of 19 mm and 16 mm for S. aureus and E. coli, respectively. To be considered as a supplement or to be used for food fortification, the fabricated ZnO must have no negative effect on gastrointestinal microbiota. Intestinal bacteria play a key role in both digestion and regulation of the immune system. For example, they are responsible for maintaining immune and metabolic homeostasis, and protecting against pathogens [13]. There are different chemical, nutritional, and immunological factors that can affect the population of human microbiota. In this regard, the properties of metallic elements such as ZnO can significantly limit gastrointestinal bacterial growth. Moreover, it is obvious that antimicrobial properties of the metallic particles can be varied by changing their physicochemical properties. In this context, the shape, particle size, and capping material are the most effective parameters [14,15]. To date, there are no reports on the successful fabrication of biocompatible ZnO particles for application as a dietary supplement. Therefore, this study was performed to synthesize a biocompatible ZnO particle using a natural heteropolysaccharide, xanthan gum (XG), as a stabilizer, thickener, and emulsifier without using any toxic chemicals. In contrast to the currently available zinc supplements, the fabrication of biocompatible micro-sized ZnO particles may be a promising Foods 2019, 8, 88 3 of 10 form of zinc supplementation, as the reduction of metal elements to a few hundreds of microns, Foods 2019, 8, x FOR PEER REVIEW 3 of 10 can substantially enhance their intestinal absorption. 2. Materials and Methods 2.1. Chemicals The zinc zinc acetate acetate dehydrate dehydrate was was obtained obtained from from Merck Merck Millipore Millipore (Darmstadt, (Darmstadt, Germany). Germany). The TheAmmonium Ammonium hydroxide, hydroxide, XG and XG andLB broths LB broths (Lysogeny (Lysogeny broth) broth) were were purchased purchased from from Sigma-Aldrich Sigma-Aldrich (St. (St.Louis, Louis, Missouri, Missouri, USA). USA). All the All solutions the solutions used usedfor the for fabrication the fabrication of nanoparticles of nanoparticles were prepared were prepared using ultrapureusing ultrapure water waterthrough through a Millipore a Millipore water water purifi purificationcation system system (Milli-Q, (Milli-Q, Milford, Milford, MA, MA, USA). USA). Ampicillin discs (10 µµg)g) and blankblank paper discs (6 mm diameter) were obtained from Fort Richard Laboratories (Auckland, New Zealand). 2.2. General Procedure for the SynthesisSynthesis of